濒危植物无距虾脊兰种子无菌萌发与幼苗形成
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摘要
[目的]揭示无距虾脊兰种子无菌萌发过程的形态特征和组织结构特点,为无距虾脊兰保育和离体快速繁殖提供参考依据。[方法]以浙江天目山无距虾脊兰野生居群为材料,采收其果实进行无菌萌发实验。通过形态观察和石蜡切片法,对种子萌发到幼苗初步形成过程进行细胞组织学研究,并对发育阶段进行划分。[结果]无距虾脊兰种子萌发的适宜基本培养基为VW培养基,椰子水对萌发具有促进作用,而香蕉泥对萌发具有抑制作用,在授粉后胚龄为4个月左右采收进行播种,萌发率最高。依据对其萌发过程中种胚膨胀、顶端分生组织的出现和发育、叶和根的分化等突出性状的观察,将无距虾脊兰种子萌发过程分为4个阶段。[结论]无距虾脊兰的种子无菌萌发过程主要由种胚吸水膨胀,原球茎的形成和分化2个生物学过程组成,其中,顶端分生组织、叶绿体和维管束为原球茎生长和发育过程中的重要结构。
[Objective] The study aims at revealing the morphological characteristics and histological structure during aseptic seed germination of Calanthe tsoongiana, so as to provide some references for its conservation and in vitro rapid propagation. [Method] This study was conducted by using wild population of C. tsoongiana distributed in Tianmushan, Zhejiang Province, aseptic germination was taken after harvesting the fruits. The process from seed germination to initial seedling formation was studied and divided into several developmental stages by morphological observation and paraffin section method. [Result] It proved that the Vacin & Went(VW) medium was the suitable basal medium for seed germination which was facilitated by coconut water. Nevertheless, the banana slurry had an inhibitory effect. The germination rate reached the highest when harvesting the fruit at the time of four months or so after pollination. The process of seed germination was divided into four stages according to the observation on swelled embryo, the presence and development of shoot apical meristem, the differentiation of young leaf and root, etc. [Conclusion] The process of aseptic seed germination is mainly composed of two biological processes, seed imbibition and the form and differentiation of protocorm. Apical meristem, chloroplast and vascular bundle are the major structures during the growth and development of protocorm.
[Objective] The study aims at revealing the morphological characteristics and histological structure during aseptic seed germination of Calanthe tsoongiana, so as to provide some references for its conservation and in vitro rapid propagation. [Method] This study was conducted by using wild population of C. tsoongiana distributed in Tianmushan, Zhejiang Province, aseptic germination was taken after harvesting the fruits. The process from seed germination to initial seedling formation was studied and divided into several developmental stages by morphological observation and paraffin section method. [Result] It proved that the Vacin & Went(VW) medium was the suitable basal medium for seed germination which was facilitated by coconut water. Nevertheless, the banana slurry had an inhibitory effect. The germination rate reached the highest when harvesting the fruit at the time of four months or so after pollination. The process of seed germination was divided into four stages according to the observation on swelled embryo, the presence and development of shoot apical meristem, the differentiation of young leaf and root, etc. [Conclusion] The process of aseptic seed germination is mainly composed of two biological processes, seed imbibition and the form and differentiation of protocorm. Apical meristem, chloroplast and vascular bundle are the major structures during the growth and development of protocorm.
引文
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[16] 胡琦敏,李勇毅,黄云峰,等.海伦兜兰的无菌播种与快速繁殖[J].植物生理学报,2016,52(9):1443-1448.
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[20] Hoang N H,Kane M E,Radcliffe E N,et al.Comparative seed germination and seedling development of the ghost orchid,Dendrophylax linden (Orchidaceae),and molecular identification of its mycorrhizal fungus from South Florida[J].Annals of Botany,2017,119(3):379-393.
[21] 聂宁,朱艳,田梅,等.白及种子萌发及原球茎发育过程的细胞组织学观察[J].中国中药杂志,2016,41(8):1446-1449.
[22] 张燕,李思锋,黎斌.独蒜兰种子无菌萌发过程观察和萌发培养基筛选[J].西北农业学报,2010,19(1):136-139.
[23] 陈进勇,程金水.几种中国兰种子试管培养根状茎发生的研究[J].北京林业大学学报,1998,20(1):35-38.
[24] Godo T,Komorli M,Nakaoki E,et al.Germination of mature seeds of Calanthe tricarinata Lindl.,an endangered terrestrial orchid,by asymbiotic culture in vitro[J].In Vitro Cellular & Developmental Biology-Plant,2010,46(3):323-328
[25] 陈汝民,叶庆生,王小菁,等.墨兰种子胚的发育和培养初步研究[J].热带亚热带植物学报,1995,3(4):72-75.
[2] 汪松,解焱.中国物种红色名录[M].北京:高等教育出版社,2004.
[3] 黄卫昌,周翔宇,倪子轶,等.基于标本和分布信息评估中国虾脊兰属植物的濒危状况[J].生物多样性,2015,23(4):493-498.
[4] 李海峰,赵志莲,张海珠,等.濒危莲瓣兰杂交育种及原生地种子萌发的研究[J].西北植物学报,2010,30(3):541-545.
[5] Kaur S,Bhutani K K.In vitro conservation and asymbiotic propagation of Coelogyne flaccida (Lindl.):a threatened orchid[J].Plant Biosyst,2014,148(5):935-944.
[6] 徐晓薇,柴明良,杨燕萍,等.寒兰杂交及种子离体萌发研究[J].园艺学报,2011,38(10):2010-2016.
[7] Edward Y.A perspective on orchid seed and protocorm development[J],Bot Stud,2017,58(1):33
[8] 连静静,钱鑫,王彩霞,等.无距虾脊兰胚珠发育及种子形成研究[J].西北植物学报,2013,32(3):494-500.
[9] Batygina T,Andronova E.The orchid protocorm:an opinion[M]// Mukerji K.Glimpses in botany.New Delhi:APH Publishing,2000:60-77.
[10] Nishimura G.Comparative morphology of Cattleya and Phalaenopsis (Orchidaceae) seedlings[J].Botanical Gazette,1981,142(3):360-365.
[11] 姚绍嫦,凌征柱,蓝祖栽,等.鹤顶兰种子萌发及原球茎增殖培养研究[J].西北植物学报,2012,32(7):1474-1479.
[12] 张燕,黎斌,李思锋.不同培养基上白芨的种子萌发与幼苗形态发生[J].西北植物学报,2009,29(8):1584-1589.
[13] Jualang A G,Devina D,Hartinie M,et al,Asymbiotic seed germination and seedling development of Vanda dearei[J].Malays Appl Biol,2014,43(2):25-33.
[14] 吴明开,李晶,沈志君,等.3种兰科植物种胚的形态发育特征观察[J].贵州农业科学,2011,39(6):19-23.
[15] 唐凤鸾,江海涛,赵志国,等,多花兰种子无菌萌发及离体快繁研究[J].广西植物,2012,32(6):793-799.
[16] 胡琦敏,李勇毅,黄云峰,等.海伦兜兰的无菌播种与快速繁殖[J].植物生理学报,2016,52(9):1443-1448.
[17] 周伟香,龚宁,李光,等.花叶开唇兰种子非共生萌发的研究[J].中草药,2007,38(4):610-613.
[18] Dutra D,Kane M E,Richardson L.Asymbiotic seed germination and in vitro seedling development of Cyrtopodium punctatum:a propagation protocol for an endangered Florida native orchid[J].Plant Cell,Tissue and Organ Culture,2009,96(3):235-243.
[19] Vudala S M,Ribas L L F.Seed storage and asymbiotic germination of Hadrolaelia grandis (Orchidaceae)[J].South African Journal of Botany,2017(1),108:1-7.
[20] Hoang N H,Kane M E,Radcliffe E N,et al.Comparative seed germination and seedling development of the ghost orchid,Dendrophylax linden (Orchidaceae),and molecular identification of its mycorrhizal fungus from South Florida[J].Annals of Botany,2017,119(3):379-393.
[21] 聂宁,朱艳,田梅,等.白及种子萌发及原球茎发育过程的细胞组织学观察[J].中国中药杂志,2016,41(8):1446-1449.
[22] 张燕,李思锋,黎斌.独蒜兰种子无菌萌发过程观察和萌发培养基筛选[J].西北农业学报,2010,19(1):136-139.
[23] 陈进勇,程金水.几种中国兰种子试管培养根状茎发生的研究[J].北京林业大学学报,1998,20(1):35-38.
[24] Godo T,Komorli M,Nakaoki E,et al.Germination of mature seeds of Calanthe tricarinata Lindl.,an endangered terrestrial orchid,by asymbiotic culture in vitro[J].In Vitro Cellular & Developmental Biology-Plant,2010,46(3):323-328
[25] 陈汝民,叶庆生,王小菁,等.墨兰种子胚的发育和培养初步研究[J].热带亚热带植物学报,1995,3(4):72-75.